Japanese encephalitis (JE) is a vector-borne zoonotic disease caused by the Japanese encephalitis virus (JEV). It causes encephalitis in human and horses, and may lead to reproductive failure in sows. The first human encephalitis case in Malaya (now Malaysia) was reported during World War II in a British prison in 1942. Later, encephalitis was observed among race horses in Singapore. In 1951, the first JEV was isolated from the brain of an encephalitis patient. The true storyline of JE exposure among humans and animals has not been documented in Malaysia. In some places such as Sarawak, JEV has been isolated from mosquitoes before an outbreak in 1992. JE is an epidemic in Malaysia except Sarawak. There are four major outbreaks reported in Pulau Langkawi (1974), Penang (1988), Perak and Negeri Sembilan (1998-1999), and Sarawak (1992). JE is considered endemic only in Sarawak. Initially, both adults and children were victims of JE in Malaysia, however, according to the current reports; JE infection is only lethal to children in Malaysia. This paper describes a timeline of JE cases (background of each case) from first detection to current status, vaccination programs against JE, diagnostic methods used in hospitals and factors which may contribute to the transmission of JE among humans and animals in Malaysia.
Traditionally, engineering and environment-based interventions have contributed to the prevention of malaria in Asia. However, with the introduction of DDT and other potent insecticides, chemical control became the dominating strategy. The renewed interest in environmental-management-based approaches for the control of malaria vectors follows the rapid development of resistance by mosquitoes to the widely used insecticides, the increasing cost of developing new chemicals, logistical constraints involved in the implementation of residual-spraying programs and the environmental concerns linked to the use of persistent organic pollutants. To guide future research and operational agendas focusing on environmental-control interventions, it is necessary to learn from the successes and failures from the time before the introduction of insecticides. The objective of this paper is to describe the experiences gained in Asia with early vector control interventions focusing on cases from the former Indian Punjab, Malaysia and Sri Lanka. The paper deals primarily with the agricultural engineering and land and water management vector control interventions implemented in the period 1900-1950. The selected cases are discussed in the wider context of environment-based approaches for the control of malaria vectors, including current relevance. Clearly, some of the interventions piloted and implemented early in the last century still have relevance today but generally in a very site-specific manner and in combination with other preventive and curative activities. Some of the approaches followed earlier on to support implementation would not be acceptable or feasible today, from a social or environmental point of view.
Cystic echinococcosis (CE) is a neglected zoonotic disease with a worldwide distribution and is a major public health problem in some areas. Diagnosis of CE is mainly based on clinical symptoms, imaging and serological testing, however, improvement in serodiagnosis is still needed. This study was aimed at detecting circulating Echinococcus antigen in CE patients using a lateral flow dipstick (LFD) assay. Three types of hydatid antigens i.e. hydatid cyst fluid (HCF), native antigen B (nAgB) and recombinant antigen B (rAgB) were prepared and polyclonal rabbit antiserum was raised against each antigen. Purified IgG fractions were prepared and a portion was conjugated to gold nanoparticles. After a series of optimizations, a final antigen detection LFD assay was developed using a combination of anti-nAgB-IgG and gold-conjugated anti-HCF-IgG. Evaluation of the assay showed that 27 out of 35 (77%) serum samples from CE patients gave positive results. Meanwhile, the test showed a diagnostic specificity of 82% when tested with sera from 38 healthy individuals and 13 patients with other parasitic diseases. In conclusion, the antigen detection LFD assay seemed to be useful for diagnosis of CE and possibly for post-treatment follow-up, and merit further evaluation studies. We foresee that it may improve serodiagnosis of CE when used in tandem with an antibody detection test.
Soft ticks (Acari: Argasidae) are the second major family of the blood feeding metastriates and vectors of a number of viral and bacterial pathogens for both humans and animals. Despite the growing effort on tick surveillance and studies worldwide, there is still limited information on the soft tick distribution in the island nations of Southeast Asia, especially species that are medically and veterinarily important. With the aim to provide an overview of the current status of knowledge on soft tick distribution in the island nations of Southeast Asia (Malaysia, Singapore, Brunei, Indonesia, the Philippines and Timor-Leste), this article reviews the species of soft ticks (Acari: Argasidae) and their associated hosts and pathogens, with the addition of a pictorial summary and list of tick species discovered in this region. The most prevalent soft tick genus is Carios, and the host species most associated with findings of soft ticks in this region are bats, particularly of the Pteropodidae and Vespertilionidae families. Furthermore, the only known pathogen originating from soft ticks in the island nations of Southeast Asia was the Keterah virus, which was isolated from Argas pusillus tick in Malaysia.
The discovery of tumour selective virus-mediated apoptosis marked the birth of an alternative cancer treatment in the form of oncolytic viruses. Even though, its oncolytic efficiency was demonstrated more than 50 years ago, safety concerns which resulted from mild to lethal side effects hampered the progress of oncolytic virus research. Since the classical oncolytic virus studies rely heavily on its natural oncolytic ability, virus manipulation was limited, thereby, restricted efforts to improve its safety. In order to circumvent such restriction, experiments involving non-human viruses such as the avian Newcastle disease virus (NDV) was conducted using cultured cells, animal models and human subjects. The corresponding reports on its significant tumour cytotoxicity along with impressive safety profile initiated immense research interest in the field of oncolytic NDV. The varying degree of oncolytic efficiency and virulency among NDV strains encouraged researchers from all around the world to experiment with their respective local NDV isolates in order to develop an oncolytic virus with desirable characteristics. Such desirable features include high tumour-killing ability, selectivity and low systemic cytotoxicity. The Malaysian field outbreak isolate, NDV strain AF2240, also currently, receives significant research attention. Apart from its high cytotoxicity against tumour cells, this strain also provided fundamental insight into NDV-mediated apoptosis mechanism which involves Bax protein recruitment as well as death receptor engagement. Studies on its ability to selectively induce apoptosis in tumour cells also resulted in a proposed p38 MAPK/NF-κB/IκBα pathway. The immunogenicity of AF2240 was also investigated through PBMC stimulation and macrophage infection. In addition, the enhanced oncolytic ability of this strain under hypoxic condition signifies its dynamic tumour tropism. This review is aimed to introduce and discuss the aforementioned details of the oncolytic AF2240 strain along with its current challenges which outlines the future research direction of this virus.
Black flies (Simuliidae) are important biting insects and vectors of diseases agents of humans and livestock. Thus, understanding the taxonomy and biodiversity of these insects is crucial for control and management of these diseases. In this study, we used mitochondrial cytochrome c oxidase I sequences to examine genetic diversity of three human-biting and possible vector black fly taxa; the Simulium asakoae species-complex, S. chamlongi and S. nigrogilvum. High levels of genetic diversity (>3.5% intraspecific genetic divergence) were found in all three taxa. Phylogenetic analyses indicated that the S. asakoae complex can be divided into seven groups with the largest group consisting of specimens from Thailand, Malaysia and Myanmar. This group most likely represents true S. asakoae. The remaining haplotypes formed groups with conspecific haplotypes or with other closely related species. Among these groups, one including S. monglaense and another including S. myanmarense suggest that certain specimens identified as S. asakoae most likely belong to those species. Therefore, they constitute new locality records for Thailand and also represent new records of anthropophily. Members of S. chamlongi are not monophyletic as its clade also included S. hackeri. A median joining network revealed strong geographic associations of the haplotypes of S. nigrogilvum suggesting limitation of gene flow. Because this species occurs mainly in high elevation habitats, low land areas could present a barrier to gene flow.
Wolbachia are maternally transmitted bacteria found in most arthropods and nematodes, but little is known about their distribution and reproductive dynamics in the Malaysian dengue vector Aedes albopictus. In this study, polymerase chain reaction (PCR) was used to determine the presence of Wolbachia from field collected Ae. albopictus from various parts of the country using wsp specific primers. Ae. albopictus had Wolbachia infection ranging from 60 to 100%. No sequence diversity of wsp gene was found within all wAlbA and wAlbB sequences. Our findings suggest that Wolbachia infection amongst the Malaysian Ae. albopictus were not homogenously distributed in all districts in Malaysia. The presence of Wolbachia in different organs of Ae. albopictus was also determined. Wolbachia were only found in the ovaries and midguts of the mosquitoes, while absent in the salivary glands. The effects of Wolbachia on Ae. albopictus fecundity, longevity and egg viability were studied using infected and uninfected colonies. The removal of Wolbachia from Ae. albopictus resulted in reduced fecundity, longevity and egg viability, thus. Wolbachia seem to play a vital role in Ae. albopictus reproductive system.
Dengue is fast becoming the most urgent health issue in Malaysia, recording close to a 10-fold increase in cases over the last decade. With much uncertainty hovering over the recently introduced tetravalent vaccine and no effective antiviral drugs, vector control remains the most important strategy in combating dengue. This study analyses the relationship between weather predictors including its lagged terms, and dengue incidence in the District of Tawau over a period of 12 years, from 2006 to 2017. A forecasting model purposed to predict future outbreaks in Tawau was then developed using this data. Monthly dengue incidence data, mean temperature, maximum temperature, minimum temperature, mean relative humidity and mean rainfall over a period of 12 years from 2006 to 2017 in Tawau were retrieved from Tawau District Health Office and the Malaysian Meteorological Department. Cross-correlation analysis between weather predictors, lagged terms of weather predictors and dengue incidences established statistically significant cross-correlation between lagged periods of weather predictors-namely maximum temperature, mean relative humidity and mean rainfall with dengue incidence at time lags of 4-6 months. These variables were then employed into 3 different methods: a multivariate Poisson regression model, a Seasonal Autoregressive Integrated Moving Average (SARIMA) model and a SARIMA with external regressors for selection. Three models were selected but the SARIMA with external regressors model utilising maximum temperature at a lag of 6 months (p-value:0.001), minimum temperature at a lag of 4 months (p-value:0.01), mean relative humidity at a lag of 2 months (p-value:0.001), and mean rainfall at a lag of 6 months (p-value:0.001) produced an AIC of 841.94, and a log-likelihood score of -413.97 establishing it as the best fitting model of the methodologies utilised. In validating the models, they were utilised to develop forecasts with the model selected with the highest accuracy of predictions being the SARIMA model predicting 1 month in advance (MAE: 7.032, MSE: 83.977). This study establishes the effect of weather on the intensity and magnitude of dengue incidence as has been previously studied. A prediction model remains a novel method of evidence-based forecasting in Tawau, Sabah. The model developed in this study, demonstrated an ability to forecast potential dengue outbreaks 1 to 4 months in advance. These findings are not dissimilar to what has been previously studied in many different countries- with temperature and humidity consistently being established as powerful predictors of dengue incidence magnitude. When used in prognostication, it can enhance- decision making and allow judicious use of resources in public health setting. Nevertheless, the model remains a work in progress- requiring larger and more diverse data.
Most female black flies in the genus Simulium are blood-sucking flies and they can cause various parasitic diseases in human and animal. A total of 94 species of black flies have been reported in Malaysia, however, their biting behavior and role as vector of infectious agents remain understudied. To fill in this knowledge gap, we attempted to survey adult black flies from field populations in Peninsular Malaysia. In a survey carried out in 2017 at Tasik Kenyir, Terengganu, three females were caught while attracted and landed on human skin. Further morphological and molecular analyses showed that the specimens were identical to Simulium (Gomphostilbia) aziruni Takaoka, Hashim & Chen of the Simulium gombakense species-group. This is the first report on a black fly species attracted to human in Malaysia which serves as a steppingstone towards in-depth studies for black flies in this region.
The larval development of Chrysomya albiceps and Lucilia sericata is a well-known and valuable tool for estimating the postmortem interval (PMI). The third instar larvae of Ch. albiceps are facultative predators of the larvae of other necrophagous dipterans, and this behaviour is undoubtedly an important factor to consider. Both species are found together during human autopsies in Spain and other parts of Europe, where they cohabit, so a precise knowledge of their larval coexistence data is essential. The aim of this study is to investigate their coexistence and the intraguild predation of Ch. albiceps on L. sericata under experimental conditions and in real case reports. To analyse intra- and interspecific competition, four densities were used under controlled abiotic conditions [25ºC, 60-70% RH and 12:12 (D:N)]. The experimental data were compared with data corresponding to their coexistence in natural conditions, so annual activity and forensic case reports for both species were studied. The results indicate that the mortality of immature specimens in both species, the preimaginal developmental time, and adult size were affected by competition. In natural conditions, adult coexistence during the spring-summer period was confirmed. The simultaneity of the two species colonising human corpses is frequent in the studied area, with Ch. albiceps being the dominant species, and L. sericata the relevant species for estimating the minimum postmortem interval.
Spotted fever group (SFG) rickettsiae causes a number of diseases in humans worldwide, which can range from mild to highly lethal. Since the clinical presentations of rickettsioses caused by SFG rickettsiae are variable and may be similar to the diseases caused by other rickettsiae, such as Orientia tsutsugamushi (agent for scrub typhus), Coxiella burnetii (agent for Q fever) and the typhus group rickettsiae (agents for epidemic and murine typhus), the accurate diagnosis of infections caused by SFG Rickettsia remains challenging especially in resource-poor settings in developing countries. This review summarizes the various diagnostic and detection tools that are currently available for the confirmation of infections by SFG rickettsiae. The advantages and challenges pertaining to the different serological and molecular detections methods, as well as new assays in development, are discussed. The utility of the detection tools contributing to the surveillance of SFG rickettsiae in arthropods and animals are reviewed.
Black flies (Diptera: Simuliidae) are known as vectors of disease agents in humans and livestock, with some species being vectors of Onchocerca volvulus, the filarial nematode that is the causative agent of human onchocerciasis. Nematode infections in adult female black flies have been reported from some areas in northern and western Thailand, but not from other regions of Thailand. In this study, wild-caught adult female black flies from the central region of Thailand were examined for infections with nematodes. Collections of adult females were carried out at Khlong Lan district, Kamphaeng Phet province, central Thailand. A molecular approach, based on the mitochondrial (cox1, 12S rRNA) and nuclear (18S rRNA) genes, was used to identify the species of nematodes recovered from the specimens collected. A total of 911 wild-caught adult black flies were collected. Simulium nigrogilvum was the most abundant species (n = 708), followed by S. doipuiense complex (n = 179), S. chamlongi (n = 11), S. umphangense (n = 10), S. chumpornense (n = 1), S. multistriatum species-group (n = 1), and S. maewongense (n = 1). Nematode infections were detected in nine specimens of S. nigrogilvum, of which two were positive for filarial worms (one worm each, infection rate 0.28%) and seven were positive for non-filarial nematodes (11 worms in total, infection rate 0.99%). The two filarial nematodes (third-stage larvae) were identified molecularly as Onchocerca species type I, while the 11 non-filarial nematodes were classified into ascaridoid (n = 2), tylenchid (n = 6) and mermithid (n = 3) nematodes. The results of this study demonstrated that adult female S. nigrogilvum were parasitized with diverse nematodes (filarial and non-filarial). Detection of the infective larvae of Onchocerca sp. type I in S. nigrogilvum confirms that occurrence of zoonotic onchocerciasis is highly possible in Thailand. Additional in-depth investigation of the morphology, life cycle and host-parasite relationship of nematodes that parasitized this black fly host is still needed.
While the East Asia Pacific (EAP) region has experienced tremendous economic growth and development, the resulting public health gains from reductions in its neglected tropical diseases (NTDs) have been less than expected due to opposing forces of urbanization, political instability, food insecurity, and climate change, together with co-morbidities with non-communicable diseases, including diabetes and hypertension. To be sure there's been progress towards the elimination of lymphatic filariasis and trachoma through mass drug administration, and there are opportunities to extend MDA to yaws and scabies, but for most of the other NTDs we'll require new biotechnologies. So far, EAP's major technology hubs in China, Japan, Malaysia, Singapore, South Korea, and Taiwan have mostly failed to shift their attention towards new innovations for the NTDs, including new drugs, diagnostics, and vaccines, and vector control. Unless this situation changes the EAP could be facing a new grim reality of unhealthy megacities beset by emerging arbovirus infections, widespread antimicrobial resistance, and urban helminth infections.
The banded krait, Bungarus fasciatus is a medically important venomous snake in Asia. The wide distribution of this species in Southeast Asia and southern China indicates potential geographical variation of the venom which may impact the clinical management of snakebite envenomation. This study investigated the intraspecific venom variation of B. fasciatus from five geographical locales through a venom decomplexing proteomic approach, followed by toxinological and immunological studies. The venom proteomes composed of a total of 9 toxin families, comprising 22 to 31 proteoforms at varying abundances. The predominant proteins were phospholipase A2 (including beta-bungarotoxin), Kunitz-type serine protease inhibitor (KSPI) and three-finger toxins (3FTx), which are toxins that cause neurotoxicity and lethality. The venom lethality varied with geographical origins of the snake, with intravenous median lethal doses (LD50) ranging from 0.45-2.55 µg/g in mice. The Thai Bungarus fasciatus monovalent antivenom (BFMAV) demonstrated a dose-dependent increasing immunological binding activity toward all venoms; however, its in vivo neutralization efficacy varied vastly with normalized potency values ranging from 3 to 28 mg/g, presumably due to the compositional differences of dominant proteins in the different venoms. The findings support that antivenom use should be optimized in different geographical areas. The development of a pan-regional antivenom may be a more sustainable solution for the treatment of snakebite envenomation.
Mitochondrial cytochrome c oxidase subunit I (COI) sequences were utilized to infer the population genetic structure of Simulium (Gomphostilbia) atratum De Meijere, an endemic simulid species to Indonesia. Both median-joining haplotype network and maximum-likelihood tree revealed two genetic lineages (A and B) within the species, with an overlap distribution in Lombok, which is situated along Wallace's line. Genetic differentiation and gene flow with varying frequencies (FST = 0.02-0.967; Nm = 0.01-10.58) were observed between populations of S. (G.) atratum, of which population pairs of different lineages showed high genetic differentiation. Notably, the high genetic distance of up to 5.92 % observed within S. (G.) atratum in Lombok was attributed to the existence of two genetically distinct lineages. The co-occurrence of distinct lineages in Lombok indicated that Wallace's line did not act as faunistic border for S. (G.) atratum in the present study. Moreover, both lineages also exhibited unimodal distributions and negative values of neutrality tests, suggesting a pattern of population expansion. The expansion and divergence time estimation suggested that the two lineages of S. (G.) atratum diverged and expanded during the Pleistocene era in Indonesia.
Data on soil-transmitted helminth (STH) infections and reinfection among Orang Asli (aborigine) schoolchildren and their nutritional and socioeconomic status were analyzed to investigate the pattern and the possible predictors of STH reinfection. In this longitudinal study, 120 (60 males and 60 females) Orang Asli primary schoolchildren aged 7-12 years and living in remote areas in Pos Betau, Kuala Lipis, Pahang were screened for the presence of STH using modified cellophane thick smear and Harada Mori techniques. The overall prevalence of ascariasis, trichuriasis and hookworm infections were 65.8, 97.5 and 10.8%, respectively. After complete deworming with a 3-day course of 400mg/daily of albendazole tablets, children were re-examined at 3 and 6 months from baseline. The reinfection rate, by one or more of STH species, at 3 months after deworming was high (49.5%) while 79.6% of the children were reinfected at 6 months after deworming. Logistic regression analyses showed that females, stunted children and those living in houses without toilets had significantly higher reinfection rates than others at 3 months (P<0.05). At 6 months, maternal employment status emerged as another predictor where children of working mothers had significantly higher reinfection rates (P=0.026). In conclusion, reinfection rate of STH is high and thus necessitates frequent and periodic deworming among children. Public health personnel need to re-look at the current control measures and identify innovative and integrated ways in order to reduce STH significantly in the rural communities.
Although several studies have reported pharmacological and immunological activity, as well as the role of black flies in transmitting pathogens to vertebrate hosts through salivary glands (SG) during blood feeding, SG proteomes of the anthropophilic black flies in Thailand have never been reported. Therefore, this study determined the SG proteomes of female S. nigrogilvum and S. nodosum. Sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) and two-dimensional (2-DE) gels containing separated SG proteins of individual species were subjected to liquid chromatography-tandem mass spectrometry (LCMS/MS) and an orthologous protein search from eukaryotic organism, nematocera and simuliidae databases for total protein identification. SDS-PAGE and protein staining revealed at least 13 and 9 major protein bands in the SGs of female S. nigrogilvum and S. nodosum, respectively, as well as several minor ones. The 2-DE demonstrated a total of 56 and 41 protein spots for S. nigrogilvum and S. nodosum, respectively. Most of the proteins obtained in both species were enzymes involved in blood feeding, including proteases, apyrases, hyaluronidases, aminopeptidase and elastase. The results obtained in this study provided a new body of knowledge for a better understanding on the role of salivary gland proteins in these black fly species in Thailand.
A parasitological survey of 16 pangolins, confiscated from the Department of Wildlife and Nature Park Peninsular Malaysia (DWNP) at Kelantan and Pulau Pinang, Malaysia was conducted in 2011. Amblyomma javanense (family: Ixodidae) was the only ectoparasite found on the pangolins. The prevalence, intensity and life cycle of A. javanense were observed together with the respective pangolins' age and sex. It was found that 68.8% of the pangolins were infected, and significant difference, χ(2)(1, N=16)=4.02, p=0.05 were observed with males higher in infestation (88.9%) as compared to the females (42.9%). However, the mean intensity was higher on females (72) as compared to males (31.6). In addition, significant difference, χ(2) (2, N=16)=6.73, p=0.05 was recorded between adults and juveniles with juveniles found to be 100% infected as compared to adult (63.6%). Nevertheless, the mean intensity was slightly higher on adults (47) than juveniles (35). Adult ticks were found in higher numbers as compared to the nymph and larvae with number of male ticks higher (236) as compared to the females (53). Similarly, a high significant difference χ(2)(2, N=469)=203.47, p=0.05 was recorded in the composition of the tick's life stages with a higher number of adult ticks (61.6%) followed by nymph (30.3%) and larvae (8.1%). As such, the results of this study revealed a picture of the A. javanense life cycle which is related to the age and gender of the Malayan Pangolin.
We aimed to systematically review evidence pertaining to the safety and efficacy of nitazoxanide in treating infectious diarrhea. On September 21, 2017, we identified relevant studies using 12 databases. The estimates of the included studies were pooled as a risk ratio (RR). We conducted a network and pairwise random-effects meta-analysis for both direct and indirect comparisons of different organisms that are known to cause diarrhea. The primary and secondary analysis outcomes were clinical response until cessation of illness, parasitological response and adverse events. We included 18 studies in our analysis. In cryptosporidiosis, the overall estimate favored nitazoxanide in its clinical response in comparison with placebo RR 1.46 [95% CI 1.22-1.74; P-value <0.0001]. Network meta-analysis among patients with Giardia intestinalis showed an increase in the probability of diarrheal cessation and parasitological responses in comparison with placebo, RR 1.69 [95% CI 1.08-2.64, P-score 0.27] and RR 2.91 [95% CI 1.72-4.91, P-score 0.55] respectively. In Clostridium difficile infection, the network meta-analysis revealed a non-significantly superior clinical response effect of nitazoxanide to metronidazole 31 days after treatment RR 1.21 [95% CI 0.87-1.69, P-score 0.26]. In Entamoeba histolytica, the overall estimate significantly favored nitazoxanide in parasitological response with placebo RR 1.80 [95% CI 1.35-2.40, P-value < 0.001]. We highlighted the effectiveness of nitazoxanide in the cessation of diarrhea caused by Cryptosporidium, Giardia intestinalis and Entamoeba histolytica infection. We also found significant superiority of NTZ to metronidazole in improving the clinical response to G. intestinalis, thus it may be a suitable candidate for treating infection-induced diarrhea. To prove the superiority of NTZ during a C. difficile infection may warrant a larger-scale clinical trial since its superiority was deemed insignificant. We recommend nitazoxanide as an appropriate option for treating infectious diarrhea.
Albendazole, a benzimidazole derivative, was administered as a single dose of either 400 mg or 600 mg to two groups to ascertain the efficacy, tolerance and safety of the regimens. At a dose of 400 mg, a cure rate 35/36 (97%) against Ascaris was found. At 600 mg, the cure rate was 21/30 (70%), significantly lower than the 400 mg rate. Against Trichuris, albendazole at 400 mg had a cure rate of 21/48 (44%), at 600 mg there was a cure rate of 29/43 (67%). Mild side effects were noted in 7 individuals. Whether the moderate increase in efficacy against Trichuris and the loss of efficacy against Ascaris improves the cost:benefit ratio must be left to the prescriber of the drug.